Programmable Self-Assembly of DNA–Protein Hybrid Hydrogel for Enzyme Encapsulation with Enhanced Biological Stability

Abstract

A DNA-protein hybrid hydrogel was constructed based on a programmable assembly approach, which served as a biomimetic physiologic matrix for efficient enzyme encapsulation. A dsDNA building block tailored with precise biotin residues was fabricated based on supersandwich hybridization, and then the addition of streptavidin triggered the formation of the DNA-protein hybrid hydrogel. The biocompatible hydrogel, which formed a flower-like porous structure that was 6.7 ± 2.1 μm in size, served as a reservoir system for enzyme encapsulation. Alcohol oxidase (AOx), which served as a representative enzyme, was encapsulated in the hybrid hydrogel using a synchronous assembly approach. The enzyme-encapsulated hydrogel was utilized to extend the duration time for ethanol removal in serum plasma and the enzyme retained 78% activity after incubation with human serum for 24 h. The DNA-protein hybrid hydrogel can mediate the intact immobilization on a streptavidin-modified and positively charged substrate, which is very beneficial to solid-phase biosensing applications. The hydrogel-encapsulated enzyme exhibited improved stability in the presence of various denaturants. For example, the encapsulated enzyme retained 60% activity after incubation at 55 °C for 30 min. The encapsulated enzyme also retains its total activity after five freeze-thaw cycles and even suspended in solution containing organic solvents.

A new paradigm in sweat based wearable diagnostics biosensors using Room Temperature Ionic Liquids (RTILs)

“…Thus enhancing the stability of affinity based capture probes is critical for the prolonged functionality and performance of wearable diagnostic biosensors 2–5 . The biochemical integrity of these capture probes need to be maintained during the continuous and prolonged exposure to sweat in order to report multiple measurements in a 24 hour period.…”

DNA Hydrogel Assemblies: Bridging Synthesis Principles to Biomedical Applications

“…Ionic interaction between DNA and sulfonium precursor of poly‐phenylenevinylene or CTAB are reported as examples of physically formed gels containing DNA backbone. In an interesting study, a hybrid DNA‐based hydrogel was fabricated via a programmable assembly approach using protein as conjugating reagent . This hierarchical hydrogel served as a biomimetic physiologic network and made based on supersandwich hybridization of dsDNA building blocks tailored with precise biotin residues, followed by the addition of streptavidin protein to the hybridization product.…”

“…A) Schematic illustration of DNA‐protein hybrid hydrogel self‐assembly. Reproduced with permission . Copyright 2016, American Chemical Society.…”

How to Construct DNA Hydrogels for Environmental Applications: Advanced Water Treatment and Environmental Analysis

“…Recently, a great progress had been made. A new DNA hydrogel took advantage of the affinity between protein and specific DNA to immobilize enzyme in the DNA‐protein porous hydrogel . The dsDNA building blocks tailored with biotin residues were triggered to format the hybrid hydrogels by the addition of streptavidin.…”